An ARL3–UNC119–RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium
- Kevin J. Wright1,
- Lisa M. Baye2,
- Anique Olivier-Mason4,
- Saikat Mukhopadhyay1,
- Liyun Sang1,
- Mandy Kwong1,
- Weiru Wang1,
- Pamela R. Pretorius2,3,
- Val C. Sheffield3,
- Piali Sengupta4,
- Diane C. Slusarski2 and
- Peter K. Jackson1,5
- 1Genentech Inc., South San Francisco, California 94080, USA;
- 2Department of Biology,
- 3Howard Hughes Medical Institute, Department of Pediatrics, University of Iowa, Iowa City, Iowa 52242, USA;
- 4Department of Biology, Brandeis University, Waltham, Massachusetts 02453, USA
Abstract
The membrane of the primary cilium is a highly specialized compartment that organizes proteins to achieve spatially ordered signaling. Disrupting ciliary organization leads to diseases called ciliopathies, with phenotypes ranging from retinal degeneration and cystic kidneys to neural tube defects. How proteins are selectively transported to and organized in the primary cilium remains unclear. Using a proteomic approach, we identified the ARL3 effector UNC119 as a binding partner of the myristoylated ciliopathy protein nephrocystin-3 (NPHP3). We mapped UNC119 binding to the N-terminal 200 residues of NPHP3 and found the interaction requires myristoylation. Creating directed mutants predicted from a structural model of the UNC119–myristate complex, we identified highly conserved phenylalanines within a hydrophobic β sandwich to be essential for myristate binding. Furthermore, we found that binding of ARL3-GTP serves to release myristoylated cargo from UNC119. Finally, we showed that ARL3, UNC119b (but not UNC119a), and the ARL3 GAP Retinitis Pigmentosa 2 (RP2) are required for NPHP3 ciliary targeting and that targeting requires UNC119b myristoyl-binding activity. Our results uncover a selective, membrane targeting GTPase cycle that delivers myristoylated proteins to the ciliary membrane and suggest that other myristoylated proteins may be similarly targeted to specialized membrane domains.
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Footnotes
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↵5 Corresponding author.
E-mail pjackson{at}gene.com.
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Supplemental material is available for this article.
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Article is online at http://www.genesdev.org/cgi/doi/10.1101/gad.173443.111.
- Received June 30, 2011.
- Accepted October 7, 2011.
- Copyright © 2011 by Cold Spring Harbor Laboratory Press